Detection of bovine group a rotavirus using rapid antigen detection kits, rt-PCR and next-generation DNA sequencing

利用快速抗原检测试剂盒、实时荧光定量PCR和新一代DNA测序技术检测牛A组轮状病毒

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作者:Fujiko Minami-Fukuda,Makoto Nagai, Hikaru Takai, Toshiaki Murakami, Tadashi Ozawa, Shinobu Tsuchiaka, Sachiko Okazaki, Yukie Katayama, Mami Oba, Naomi Nishiura, Yukiko Sassa, Tsutomu Omatsu, Tetsuya Furuya, Satoshi Koyama, Junsuke Shirai, Hiroshi Tsunemitsu, Yoshiki Fujii, Kazuhiko Katayama, Tetsuya Mizutani

Abstract

We investigated the sensitivity of human rotavirus rapid antigen detection (RAD) kits, RT-PCR and next-generation DNA sequencing (NGS) for detection of bovine group A rotavirus (RVA). The Dipstick 'Eiken' Rota (Dipstick) showed the highest sensitivity out of the seven RAD kits against all selected strains in limited dilution analyses, which was consistent with the results for equine rotavirus previously reported. RT-PCR had 10⁰-10³-fold higher sensitivity than the Dipstick. NGS using thirteen RT-PCR-negative fecal samples revealed that all samples yielded RVA reads and especially that two of them covered all 11 genome segments. Moreover, mapping reads to reference sequences allowed genotyping. The NGS would be sensitive and useful for analysis of less dependent on specific primers and screening of genotypes.

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